Culture and identification of dendritic cells from mouse bone marrow in vitro

Abstract

Establish a method of cultivation and purification of dendritic cells (DCs) from mouse bone marrow in vitro to provide experimental material and establish basis for the study of immunological tolerance. BALB/c (H-2d) mice spleen lymphocytes and bone marrow cells were extracted under sterile conditions and cultured in RPMI 1640 medium. DCs were induced by recombinant granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin (IL-4). Then, the morphology of DCs was observed under light microscopy and CD80, CD86 expression levels were detected by flow cytometry. The results showed that bone marrow cells appeared irregular and formed dendritic processes after 3 days cultured with stimulating factors in vitro which present the typical morphology of dendritic cells; Costimulatory molecules (CD80, CD86) were low expression on the cellular surface. Obtaining DCs from bone marrow cells is a promising method, which might meet further study on function, marker and anti-tumor role of DCs in vitro.